磷酸鞘胺醇受体在四氯化碳引起的肝纤维化小鼠肝脏的表达

Expression of Sphingosine 1-phosphate Receptors in Fibrotic Liver Induced by Carbon Tetrachloride

目的研究四氯化碳(CCl4)引起的肝纤维化小鼠肝组织中的磷酸鞘胺醇(sphingosine 1-phosphate,S1P)受体1、2、3(S1P1-3)mRNA和蛋白含量的变化。方法制备小鼠CCl4肝纤维化模型,采用实时荧光定量聚合酶链反应方法测定小鼠肝组织S1P1-3mRNA表达情况;采用免疫印记(Western blotting)方法测定小鼠肝组织S1P1-3蛋白表达情况。结果CCl4诱导小鼠肝纤维化2周或4周后,与对照组相比,小鼠肝组织的S1P1mRNA的表达差异无统计学意义,而S1P2、S1P3mRNA的表达显著上调(P<0.05);与之相对应,S1P1的蛋白表达差异无统计学意义,而S1P2、S1P3的蛋白表达明显增加(P<0.05)。结论在CCl4引起的小鼠肝纤维化形成过程中,S1P、S1P在mRNA和蛋白水平均显著上调,表明S1P相关受体在肝纤维化发生过程中起重要作用。

Objective To determine the mRNA and protein expression of sphingosine 1-phosphate receptors 1,2 and 3（S1P1-3） in the normal and fibrotic liver.Methods ICR mice were randomized into two experimental groups,control [olive oil（OO） treated] and model（CCl4 treated） groups.Mice were sacrificed at two or four weeks of treatment.Liver tissue was examined by real-time RT-PCR and Western blotting to analyze the expression of S1P1-3.Results Two or four weeks treatment by CCl4,S1P1 mRNA expression was not significantly different;S1P2,S1P3 mRNA expression was up-regulated（P0.05） in liver tissue.Correspondingly,S1P1 protein level was not significantly different;S1P2,S1P3 protein level was up-regulated（P0.05）.Conclusion During the process of liver fibrosis,sphingosine 1-phosphate receptor1-3 mRNA expression and protein level were various,the relative mRNA and protein level expression of S1P2,S1P3 have shown increases.Our results indicate that S1P receptors have an extremely important role in liver fibrogenesis.